Accessing and integrating data and knowledge for biomedical research.
To review the issues that have arisen with the advent of translational research in terms of integration of data and knowledge, and survey current efforts to address these issues. Using examples form the biomedical literature, we identified new trends in biomedical research and their impact on bioinformatics. We analyzed the requirements for effective knowledge repositories and studied issues in the integration of biomedical knowledge. New diagnostic and therapeutic approaches based on gene expression patterns have brought about new issues in the statistical analysis of data, and new workflows are needed are needed to support translational research. Interoperable data repositories based on standard annotations, infrastructures and services are needed to support the pooling and meta-analysis of data, as well as their comparison to earlier experiments. High-quality, integrated ontologies and knowledge bases serve as a source of prior knowledge used in combination with traditional data mining techniques and contribute to the development of more effective data analysis strategies. As biomedical research evolves from traditional clinical and biological investigations towards omics sciences and translational research, specific needs have emerged, including integrating data collected in research studies with patient clinical data, linking omics knowledge with medical knowledge, modeling the molecular basis of diseases, and developing tools that support in-depth analysis of research data. As such, translational research illustrates the need to bridge the gap between bioinformatics and medical informatics, and opens new avenues for biomedical informatics research.


CNS immune surveillance and neuroinflammation: endocannabinoids keep control.
To avoid inflammatory escalation, the central nervous system (CNS) harbors an impressive arsenal of cellular and molecular mechanisms enabling strict control of immune reactions. We here summarize studies suggesting that the old paradigm of the "CNS immune privilege" is overly simplistic. The immune system is allowed to keep the CNS under surveillance, but in a strictly controlled, limited and well-regulated manner. The first line of defense lies outside the brain parenchyma to spare neuronal tissue from the detrimental effects of an inflammatory immune response. As a second line of defense neuroinflammation is unavoidable when pathogens infiltrate the brain or the CNS-immune-homeostasis fails. Inflammation in the CNS is often accompanied by divers brain pathologies. We here review recent strategies to maintain brain homeostasis and modulate neuroinflammation. We focus on Multiple Sclerosis as an example of a complex neuroinflammatory disease. In the past years, several in vitro, in vivo and clinical studies suggested that the endocannabinoid system participates crucially in the immune control and protection of the CNS. We discuss here the endocannabinoid system as a key regulator mechanism of the cross talk between brain and the immune system as well as its potential as a therapeutic target.


A new auroral phenomenon, the anti-black aurora.
Black auroras are small-scale features embedded in the diffuse background aurora, typically occurring post-substorm after magnetic midnight and with an eastward drift imposed. Black auroras show a significant reduction in optical brightness compared to the surrounding diffuse aurora, and can appear as slow-moving arcs or rapidly-moving patches and arc segments. We report, for the first time, an even more elusive small-scale optical structure that has always been observed occurring paired with [Formula: see text] 10% of black aurora patches. A patch or arc segment of enhanced luminosity, distinctly brighter than the diffuse background, which we name the anti-black aurora, may appear adjacent to the black aurora. The anti-black aurora is of similar shape and size, and always moves in parallel to the drifting black aurora, although it may suddenly switch sides for no apparent reason. The paired phenomenon always drifts with the same average speed in an easterly direction. From the first dual-wavelength (427.8 nm and 844.6 nm) optical observations of the phenomenon recorded on 12 March 2016 outside Tromsø Norway, we show that the anti-black and black auroras have a higher and lower mean energy, respectively, of the precipitating electrons compared to the diffuse background.


H5N1 highly pathogenic avian influenza in Southeast Asia.
Since the first H5N1 highly pathogenic avian influenza virus (HPAIV) infection in the region in August 2003, Cambodia, Laos, Malaysia, Myanmar, Indonesia, Thailand and Vietnam have all recorded outbreaks of the disease. The HPAIV continues to occur in some countries in Southeast Asia despite control programmes encompassing surveillance, vaccination and stamping out strategies. A number of strains have been circulating in the region since the first outbreaks in 2003, and although the source of the initial outbreaks in domestic poultry is not known, the continuing propagation of disease in the region is primarily the result of the movement of domestic poultry and poultry products, and people. A comprehensive approach using all the strategies available to break the chain of transmission of the virus in poultry will be needed to achieve lasting disease control.


Tetrahydrobiopterin and alkylglycerol monooxygenase substantially alter the murine macrophage lipidome.
Tetrahydrobiopterin is a cofactor synthesized from GTP with well-known roles in enzymatic nitric oxide synthesis and aromatic amino acid hydroxylation. It is used to treat mild forms of phenylketonuria. Less is known about the role of tetrahydrobiopterin in lipid metabolism, although it is essential for irreversible ether lipid cleavage by alkylglycerol monooxygenase. Here we found intracellular alkylglycerol monooxygenase activity to be an important regulator of alkylglycerol metabolism in intact murine RAW264.7 macrophage-like cells. Alkylglycerol monooxygenase was expressed and active also in primary mouse bone marrow-derived monocytes and "alternatively activated" M2 macrophages obtained by interleukin 4 treatment, but almost missing in M1 macrophages obtained by IFN-γ and lipopolysaccharide treatment. The cellular lipidome of RAW264.7 was markedly changed in a parallel way by modulation of alkylglycerol monooxygenase expression and of tetrahydrobiopterin biosynthesis affecting not only various ether lipid species upstream of alkylglycerol monooxygenase but also other more complex lipids including glycosylated ceramides and cardiolipins, which have no direct connection to ether lipid pathways. Alkylglycerol monooxygenase activity manipulation modulated the IFN-γ/lipopolysaccharide-induced expression of inducible nitric oxide synthase, interleukin-1β, and interleukin 1 receptor antagonist but not transforming growth factor β1, suggesting that alkylglycerol monooxygenase activity affects IFN-γ/lipopolysaccharide signaling. Our results demonstrate a central role of tetrahydrobiopterin and alkylglycerol monooxygenase in ether lipid metabolism of murine macrophages and reveal that alteration of alkylglycerol monooxygenase activity has a profound impact on the lipidome also beyond the class of ether lipids.


Mechanical contribution of endocardium during finite extension and torsion experiments on papillary muscles.
Finite extension and torsion tests on cardiac papillary muscles are presently the best way to directly measure the response to shear along myocardial fibers. Quantifying this response is necessary for determining the complete three-dimensional constitutive behavior of myocardium as a transversely isotropic material. Analysis of such tests is complicated, however, since papillary muscles are materially inhomogeneous, consisting of a myocardial core surrounded by an endocardial sheath that is rich in collagen. In this article, we show that the papillary muscle response to extension and torsion additively decouples into the response of the bare myocardial core plus the response of an endocardial sheath filled with fluid (assuming the muscle is a radially inhomogeneous and incompressible continuum with cylindrical symmetry). This result allows the endocardial response to be subtracted from the intact papillary muscle response to obtain the response of the bare myocardial core. An initial estimate suggests that the endocardial sheath affects the axial moment significantly (50% of torque for all twists at low stretch) but affects the axial force only slightly (<10% at moderate twists).


Mechanisms by which nephrectomy stimulates adrenal renin.
Renin has been identified in the adrenal gland by several investigators. Nephrectomy is the most potent stimulator of adrenal renin, and in the present study we investigated the mechanism by which nephrectomy stimulates adrenal renin. The pituitary plays a permissive role since hypophysectomy abolished the response of adrenal renin to nephrectomy (from 117.3 +/- 14.55 to 10.37 +/- 1.63 ng angiotensin I/mg protein/hr) and adrenocorticotropic hormone (ACTH) treatment restored the response to nephrectomy in hypophysectomized rats to 120 +/- 20.62 ng angiotensin I/mg protein/hr. However, large doses of ACTH given to intact rats did not increase adrenal renin to the high level observed after nephrectomy. Potassium also plays an important role, since prevention of hyperkalemia after nephrectomy by treatment with a cation exchange resin, sodium polystyrene sulfonate (Kayexalate), significantly reduced the adrenal renin response to nephrectomy. A third factor involved is the lack of negative feedback by plasma angiotensin II. Infusion of angiotensin II intraperitoneally prevented the rise in adrenal renin after nephrectomy (from 65.25 +/- 7.60 to 9.27 +/- 0.99 ng angiotensin I/mg protein/hr) despite an increase in plasma potassium and corticosterone. In conclusion, three factors influence the response of adrenal renin to nephrectomy: 1) the pituitary through the release of ACTH, 2) a direct stimulation by high plasma potassium levels, 3) the lack of angiotensin II feedback inhibition. Whether the high adrenal renin contributes to the high aldosterone observed in rats after nephrectomy remains to be established.


TWIK-1 two-pore domain potassium channels change ion selectivity and conduct inward leak sodium currents in hypokalemia.
Background potassium (K+) channels, which are normally selectively permeable to K+, maintain the cardiac resting membrane potential at around -80 mV. In subphysiological extracellular K+ concentrations ([K+]o), which occur in pathological hypokalemia, the resting membrane potential of human cardiomyocytes can depolarize to around -50 mV, whereas rat and mouse cardiomyocytes become hyperpolarized, consistent with the Nernst equation for K+. This paradoxical depolarization of cardiomyocytes in subphysiological [K+]o, which may contribute to cardiac arrhythmias, is thought to involve an inward leak sodium (Na+) current. Here, we show that human cardiac TWIK-1 (also known as K2P1) two-pore domain K+ channels change ion selectivity, becoming permeable to external Na+, and conduct inward leak Na+ currents in subphysiological [K+]o. A specific threonine residue (Thr118) within the pore selectivity sequence TxGYG was required for this altered ion selectivity. Mouse cardiomyocyte-derived HL-1 cells exhibited paradoxical depolarization with ectopic expression of TWIK-1 channels, whereas TWIK-1 knockdown in human spherical primary cardiac myocytes eliminated paradoxical depolarization. These findings indicate that ion selectivity of TWIK-1 K+ channels changes during pathological hypokalemia, elucidate a molecular basis for inward leak Na+ currents that could trigger or contribute to cardiac paradoxical depolarization in lowered [K+]o, and identify a mechanism for regulating cardiac excitability.


Circadian rhythm of transferrin receptor 1 gene expression controlled by c-Myc in colon cancer-bearing mice.
The abundance of cell surface levels of transferrin receptor 1 (TfR1), which regulates the uptake of iron-bound transferring, correlates with the rate of cell proliferation. Because TfR1 expression is higher in cancer cells than in normal cells, it offers a target for cancer therapy. In this study, we found that the expression of TfR1 in mouse colon cancer cells was affected by the circadian organization of the molecular clock. The core circadian oscillator is composed of an autoregulatory transcription-translation feedback loop, in which CLOCK and BMAL1 are positive regulators and the Period (Per), Cryptochrome (Cry), and Dec genes act as negative regulators. TfR1 in colon cancer-bearing mice exhibited a 24-hour rhythm in mRNA and protein levels. Luciferase reporter analysis and chromatin immunoprecipitation experiments suggested that the clock-controlled gene c-MYC rhythmically activated the transcription of the TfR1 gene. Platinum incorporation into tumor DNA and the antitumor efficacy of transferrin-conjugated liposome-delivered oxaliplatin could be enhanced by drug administration at times when TfR1 expression increased. Our findings suggest that the 24-hour rhythm of TfR1 expression may form an important aspect of strategies for TfR1-targeted cancer therapy.


OptiFlex: Multi-Frame Animal Pose Estimation Combining Deep Learning With Optical Flow.
Animal pose estimation tools based on deep learning have greatly improved animal behaviour quantification. These tools perform pose estimation on individual video frames, but do not account for variability of animal body shape in their prediction and evaluation. Here, we introduce a novel multi-frame animal pose estimation framework, referred to as OptiFlex. This framework integrates a flexible base model (i.e., FlexibleBaseline), which accounts for variability in animal body shape, with an OpticalFlow model that incorporates temporal context from nearby video frames. Pose estimation can be optimised using multi-view information to leverage all four dimensions (3D space and time). We evaluate FlexibleBaseline using datasets of four different lab animal species (mouse, fruit fly, zebrafish, and monkey) and introduce an intuitive evaluation metric-adjusted percentage of correct key points (aPCK). Our analyses show that OptiFlex provides prediction accuracy that outperforms current deep learning based tools, highlighting its potential for studying a wide range of behaviours across different animal species.


NFIL3 is a regulator of IL-12 p40 in macrophages and mucosal immunity.
Regulation of innate inflammatory responses against the enteric microbiota is essential for the maintenance of intestinal homeostasis. Key participants in innate defenses are macrophages. In these studies, the basic leucine zipper protein, NFIL3, is identified as a regulatory transcription factor in macrophages, controlling IL-12 p40 production induced by bacterial products and the enteric microbiota. Exposure to commensal bacteria and bacterial products induced NFIL3 in cultured macrophages and in vivo. The Il12b promoter has a putative DNA-binding element for NFIL3. Basal and LPS-activated NFIL3 binding to this site was confirmed by chromatin immunoprecipitation. LPS-induced Il12b promoter activity was inhibited by NFIL3 expression and augmented by NFIL3-short hairpin RNA in an Il12b-bacterial artificial chromosome-GFP reporter macrophage line. Il12b inhibition by NFIL3 does not require IL-10 expression, but a C-terminal minimal repression domain is necessary. Furthermore, colonic CD11b(+) lamina propria mononuclear cells from Nfil3(-/-) mice spontaneously expressed Il12b mRNA. Importantly, lower expression of NFIL3 was observed in CD14(+) lamina propria mononuclear cells from Crohn's disease and ulcerative colitis patients compared with control subjects. Likewise, no induction of Nfil3 was observed in colons of colitis-prone Il10(-/-) mice transitioned from germ-free to a conventional microbiota. In conclusion, these experiments characterize NFIL3 as an Il12b transcriptional inhibitor. Interactions of macrophages with the enteric microbiota induce NFIL3 to limit their inflammatory capacity. Furthermore, altered intestinal NFIL3 expression may have implications for the pathogenesis of experimental and human inflammatory bowel diseases.


Nonconvergence in logistic and poisson models for neural spiking.
Generalized linear models are an increasingly common approach for spike train data analysis. For the logistic and Poisson models, one possible difficulty is that iterative algorithms for computing parameter estimates may not converge because of certain data configurations. For the logistic model, these configurations are called complete and quasi-complete separation. We show that these features are likely to occur because of refractory periods of neurons. We use an example to study how standard software deals with this difficulty. For the Poisson model, we show that the same difficulties arise, this time possibly due to bursting or specifics of the binning. We characterize the nonconvergent configurations for both models, show that they can be detected by linear programming methods, and discuss possible remedies.


The contributions of sex and drinking history to the CT brain scan changes in alcoholics.
Consecutive series of male and female alcoholics, Alcoholic Anonymous (AA) members and controls were examined by interview and with a CT brain scan. Male controls were shown to have larger CT brain parameters than healthy females of the same age. Female alcoholics revealed equivalent CT scan abnormalities, apart from less sulcal widening, after a markedly shorter drinking history and at a lower estimated peak alcohol consumption than male alcoholics. The CT scan findings persisted after accounting for body weight and after matching for age and length of drinking history. The CT scan parameters of female AA members approached control values more completely and after briefer abstinence than did those of male AA members. Methodological problems and sex differences in selection and other processes are discussed. The findings are consistent with sex differences in the vulnerability of the brain to alcohol toxicity, and in its recovery with abstinence.


The Cutaneous Assessment Tool: development and reliability in juvenile idiopathic inflammatory myopathy.
Clinical care and therapeutic trials in idiopathic inflammatory myopathies (IIM) require accurate and consistent assessment of cutaneous involvement. The Cutaneous Assessment Tool (CAT) was designed to measure skin activity and damage in IIM. We describe the development and inter-rater reliability of the CAT, and the frequency of lesions endorsed in a large population of juvenile IIM patients. The CAT includes 10 activity, 4 damage and 7 combined lesions. Thirty-two photographic slides depicting IIM skin lesions were assessed by 11 raters. One hundred and twenty-three children were assessed by 11 paediatric rheumatologists at 10 centres. Inter-rater reliability was assessed using simple agreements and intra-class correlation coefficients (ICC). Simple agreements in recognizing lesions as present or absent were generally high (0.5-1.0). ICCs for CAT lesions were moderate (0.4-0.75) in both slides and real patients. ICCs for the CAT activity and damage scores were 0.71 and 0.81, respectively. CAT activity scores ranged from 0 to 44 (median 7, potential range 0-96) and CAT damage scores ranged from 0 to 13 (median 1, potential range 0-22). The most common cutaneous lesions endorsed were periungual capillary loop changes (63%), Gottron's papules/sign (53%), heliotrope rash (49%) and malar/facial erythema (49%). Total CAT activity and damage scores have moderate to good reliability. Assessors generally agree on the presence of a variety of cutaneous lesions. The CAT is a promising, semi-quantitative tool to comprehensively assess skin disease activity and damage in IIM.


Homopolymeric tracts represent a general regulatory mechanism in prokaryotes.
While, traditionally, regulation of gene expression can be grouped into transcriptional, translational, and post-translational mechanisms, some mechanisms of rapid genetic variation can also contribute to regulation of gene expression, e.g., phase variation. We show here that prokaryotes evolved to include homopolymeric tracts (HTs) within coding genes as a system that allows for efficient gene inactivation. Analyses of 81 bacterial and 18 archaeal genomes showed that poly(A) and poly(T) HTs are overrepresented in these genomes and preferentially located at the 5' end of coding genes. Location of HTs at the 5' end is not driven by a preferential placement of aminoacids encoded by the AAA and TTT codons at the N-terminal of proteins. The inlA gene of the pathogen L. monocytogenes was used as a model to further study the role of HTs in reversible gene inactivation. In a number of L. monocytogenes strains, inlA harbors a 5' poly(A) HT, which regularly shows frameshift mutation leading to expression of a truncated 8 aa InlA protein. Translational fusions of the inlA 5' end allowed us to estimate that the frequency of variation in this HT is about 1,000 fold higher than the estimated average point mutation frequency. As frameshift mutations in HTs can occur at high frequencies and enable efficient gene inactivation, hypermutable HTs appear to represent a universal system for regulation of gene expression in prokaryotes. Combined with other studies indicating that HTs also enable rapid diversification of both coding and regulatory genetic sequences in eukaryotes, our data suggest that hypermutable HTs represent a general and rapid evolutionary mechanism facilitating adaptation and gene regulation across diverse organisms.


Development and use of a transition readiness scale to help manage ACT team capacity.
This article describes the creation, validation, and use of an assertive community treatment (ACT) Transition Readiness Scale (TRS) to identify clients who may be ready to transition from ACT services. Scale development was prompted by concerns over long stays on ACT teams and the resulting impact on access. Data were extracted from a centralized clinical reporting system for all 1,365 persons enrolled for at least 12 months as of August 2008 in the 42 ACT teams in New York City, including 382 clients of eight of those ACT teams. Data in seven domains deemed relevant to transition readiness were used to calculate readiness scores for each client. An algorithm assigned clients to one of three categories: consider for transition, readiness unclear, and not ready. Via the TRS algorithm, of the 1,365 clients, 192 (14%) were assigned to the consider-for-transition group, 382 (28%) to the unclear group, and 791 (58%) to the not ready group. Clinicians on the eight ACT teams categorized 15% of their current clients in the consider-for-transition group, whereas the TRS algorithm classified 18% in this category. Overall, the TRS agreed with the category assigned by ACT team clinicians in 69% of cases. The TRS may provide ACT teams and program administrators with a tool to identify clients who may be ready to transition to less intensive services, thereby opening scarce slots. Because ACT cases are complex, data summaries can offer useful syntheses of information, particularly when data from several assessments are used to summarize a client's trajectory.


Intravenous fluids in traumatic brain injury: what's the solution?
Intravenous fluid is a fundamental component of trauma care and fluid management influences patient outcomes. This narrative review appraises recent clinical studies of fluid therapy in patients with traumatic brain injury (TBI), with respect to its use in volume resuscitation and prevention of secondary injury. Despite the development of level 1 evidence in fluid resuscitation, in patients with TBI key questions concerning optimal composition and volume remain unanswered. In the absence of randomized trials demonstrating patient outcome differences, clinical practice is often based on physiological principles and surrogate endpoints. There is a physiological rationale why excessive fluid administration and positive fluid balance may increase brain swelling and intracranial pressure (ICP); in some patients, a lower cumulative fluid balance may improve outcomes, but limited human data exist. Resuscitation with 4% albumin in TBI patients in ICU worsens mortality, which may be mediated by increased ICP during the first week after injury. Hypertonic saline and mannitol decrease ICP, but may not improve survival or neurological outcomes. Sodium lactate may be a future therapy for treatment and prevention of secondary brain injury. In patients with TBI, intravenous fluids are integral to management; they may be both a source of harm and a potential therapy to limit secondary brain injury. They should be prescribed in accordance with other pharmaceutical or therapeutic interventions. Refined usage may improve patient outcomes.


Whole genome-based population biology and epidemiological surveillance of Listeria monocytogenes.
Listeria monocytogenes (Lm) is a major human foodborne pathogen. Numerous Lm outbreaks have been reported worldwide and associated with a high case fatality rate, reinforcing the need for strongly coordinated surveillance and outbreak control. We developed a universally applicable genome-wide strain genotyping approach and investigated the population diversity of Lm using 1,696 isolates from diverse sources and geographical locations. We define, with unprecedented precision, the population structure of Lm, demonstrate the occurrence of international circulation of strains and reveal the extent of heterogeneity in virulence and stress resistance genomic features among clinical and food isolates. Using historical isolates, we show that the evolutionary rate of Lm from lineage I and lineage II is low (∼2.5 × 10-7 substitutions per site per year, as inferred from the core genome) and that major sublineages (corresponding to so-called 'epidemic clones') are estimated to be at least 50-150 years old. This work demonstrates the urgent need to monitor Lm strains at the global level and provides the unified approach needed for global harmonization of Lm genome-based typing and population biology.


Effects of sexual dimorphism and landscape composition on the trophic behavior of Greater Prairie-Chicken.
Partitioning of ecological niche is expected in lekking species that show marked sexual size dimorphism as a consequence of sex-specific ecological constraints. However, niche partitioning is uncertain in species with moderate sexual dimorphism. In addition, the ecological niche of a species may also be affected by landscape composition; particularly, agricultural fragmentation may greatly influence the trophic behavior of herbivores. We studied trophic niche variation in Greater Prairie-Chickens (Tympanuchus cupido), a grouse species that shows moderate sex-dimorphism. Greater Prairie-Chickens are native to tallgrass prairies of North America, although populations persist in less natural mosaics of cropland and native habitats. We used stable isotope analysis of carbon and nitrogen in blood, claws and feathers to assess seasonal differences in trophic niche breadth and individual specialization between male and female Greater Prairie-Chickens, and between birds living in continuous and fragmented landscapes. We found that females showed broader niches and higher individual specialization than males, especially in winter and autumn. However, differences between females and males were smaller in spring when birds converge at leks, suggesting that females and males may exhibit similar feeding behaviors during the lekking period. In addition, we found that birds living in native prairies showed greater annual trophic variability than conspecifics in agricultural mosaic landscapes. Native habitats may provide greater dietary diversity, resulting in greater diversity of feeding strategies.


Commercial whey products promote intestinal barrier function with glycomacropeptide enhanced activity in downregulating bacterial endotoxin lipopolysaccharides (LPS)-induced inflammation in vitro.
Cheese whey contains bioactive compounds which have shown multiple health-promoting benefits. This study aimed to assess the commercial whey products (CWP) whey protein isolate (WPI), galacto-oligosaccharide-whey protein concentrate (GOS-W) and glycomacropeptide (GMP) for their potential to improve intestinal health in vitro using HT29-MTX intestinal goblet and Caco-2 epithelial cells. Results from HT29-MTX culture showed that WPI mitigated reactive oxygen species (ROS) production at a higher extent compared to GOS-W or GMP. However, GMP downregulated the lipopolysaccharide (LPS)-induced TLR-4 inflammatory pathway with the highest potency compared to the other CWP. Biomarkers of epithelial integrity assessed on both cell lines showed tight junction proteins claudin-1, claudin-3, occludin (OCC), and zonula occludens-1 (ZO-1) upregulation by GMP in HT29-MTX (1.33-1.93-fold of control) and in Caco-2 cells (1.56-2.09-fold of control). All CWP increased transepithelial electrical resistance (TEER) in TNF-α challenged Caco-2/HT29-MTX co-culture monolayer (p < 0.05), but only GMP was similar to the positive control TGF-β1, known for its role in promoting epithelial barrier function. The TNF-α-induced co-culture monolayer permeability was prevented at similar levels by all CWP (p < 0.05). In conclusion, CWP may be used as functional food ingredients to protect against intestinal disorders with emphasis on the GMP enhanced anti-inflammatory and intestinal barrier function properties. Further in vivo studies are guaranteed to validate these findings.


Avian and Mammalian mitochondrial ammonia-detoxifying systems in tortoise liver.
Liver mitochondria from the desert tortoise Gopherus agassizii and the Texas tortoise G. berlandieri contain glutamine synthetase, the enzyme used by birds and higher reptiles to detoxify ammonia generated intramitochondrially during amino acid catabolism. Tortoise mitochondria also contain carbamyl phosphate synthetase-I and ornithine transcarbamylase, the enzymes used by mammals for this purpose. Since chelonid reptiles have changed little since their divergence from the stem reptiles, this finding suggests that both systems were present in the latter before the emergence of the ruling reptile, avian, and mammalian lines of descent.


How Much Should We Pay to Minimize Pancreatic Leak? The Cost-effectiveness of Pasireotide in Pancreatic Resection: RETRACTED.
Pasireotide was recently shown to decrease leak rates after pancreatic resection, though the significant cost of the drug may be prohibitive. We conducted a cost-effectiveness analysis to determine whether prophylactic pasireotide possesses a reasonable cost profile by improving outcomes. A cost-effectiveness model was constructed to compare pasireotide administration after pancreatic resection versus usual care, populated by probabilities of clinical outcomes from a recent randomized trial and hospital costs (2013 US$) from a university pancreatic disease center. Sensitivity analyses were performed to identify the most influential clinical components of the model. Without considering pasireotide cost, prophylactic use of the drug saved an average of $8,109 per patient. However, when the cost of pasireotide was included, per patient costs increased from $42,159 to $77,202. This was associated with a 56% reduction in pancreatic fistula/pancreatic leak/abscess (PF/PL/A) (21.9% to 9.2%). The resultant cost per PF/PL/A avoided was $301,628. Threshold analysis demonstrated that for this intervention to be cost neutral, either the purchase price of pasireotide ($43,172) must be reduced by 92.3% (to $3324) or drug reimbursement must be $39,848. Sensitivity analyses exploring variable perioperative mortality, rate of PF/PL/A, and readmission rates did not significantly alter model outcomes. Our analyses demonstrate that when prophylactic pasireotide is administered, the cost per PF/PL/A avoided is approximately $300,000. Aggressive pricing negotiation, payer reimbursement for the drug, high-volume use, and consensus among the public, payers, and surgical community regarding the value of reducing morbidity will ultimately determine the utility of widespread pasireotide application in pancreatic resection.


Adenylate cycalse toxin of Bordetella pertussis inhibits TLR-induced IRF-1 and IRF-8 activation and IL-12 production and enhances IL-10 through MAPK activation in dendritic cells.
Adenylate cyclase toxin (CyaA) of Bordetella pertussis binds to CD11b/CD18 on macrophages and dendritic cells (DC) and confers virulence to the bacteria by subverting innate immune responses of the host. We have previously demonstrated that CyaA promotes the induction of IL-10-secreting regulatory T cells in vivo by modulating DC activation. Here, we examine the mechanism of immune subversion, specifically, the modulation of TLR signaling pathways in DC. We found that CyaA synergized with LPS to induce IL-10 mRNA and protein expression in DC but significantly inhibited IL-12p70 production. CyaA enhanced LPS-induced phosphorylation of p38 MAPK and ERK in DC, and inhibitors of p38 MAPK, MEK, or NF-kappaB suppressed IL-10 production in response to LPS and CyaA. However, inhibition of p38 MAPK, MEK, and NF-kappaB did not reverse the inhibitory effect of CyaA on TLR agonist-induced IL-12 production. Furthermore, CyaA suppression of IL-12 was independent of IL-10. In contrast, CyaA suppressed LPS- and IFN-gamma-induced IFN-regulatory factor-1 (IRF-1) and IRF-8 expression in DC. The modulatory effects of CyaA were dependent on adenylate cyclase activity and induction of intracellular cAMP, as an enzyme-inactive mutant of CyaA failed to modulate TLR-induced signaling in DC, whereas the effects of the wild-type toxin were mimicked by stimulation of the DC with PGE2. Our findings demonstrate that CyaA modulates TLR agonist-induced IL-10 and IL-12p70 production in DC by, respectively, enhancing MAPK phosphorylation and inhibiting IRF-1 and IRF-8 expression and that this is mediated by elevation of intercellular cAMP concentrations.


Ethical brain stimulation - neuroethics of deep brain stimulation in research and clinical practice.
Deep brain stimulation (DBS) is a clinically established procedure for treating severe motor symptoms in patients suffering from end-stage Parkinson's disease, dystonia and essential tremor. Currently, it is tested for further indications including psychiatric disorders like major depression and a variety of other diseases. However, ethical issues of DBS demand continuing discussion. Analysing neuroethical and clinical literature, five major topics concerning the ethics of DBS in clinical practice were identified: thorough examination and weighing of risks and benefits; selecting patients fairly; protecting the health of children in paediatric DBS; special issues concerning patients' autonomy; and the normative impact of quality of life measurements. In exploring DBS for further applications, additionally, issues of research ethics have to be considered. Of special importance in this context are questions such as what additional value is generated by the research, how to realise scientific validity, which patients should be included, and how to achieve an acceptable risk-benefit ratio. Patients' benefit is central for ethical evaluation. This criterion can outweigh very serious side-effects, and can make DBS appropriate even in paediatrics. Because standard test procedures evade central aspects of patients' benefits, measuring quality of life should be supplemented by open in-depth interviews to provide a more adequate picture of patients' post-surgical situation. To examine its entire therapeutic potential, further research in DBS is needed. Studies should be based on solid scientific hypotheses and proceed cautiously to benefit severely suffering patients without putting them to undue risks.


Fructose, high-fructose corn syrup, sucrose, and nonalcoholic fatty liver disease or indexes of liver health: a systematic review and meta-analysis.
Concerns have been raised about the concurrent temporal trend between simple sugar intakes, especially of fructose or high-fructose corn syrup (HFCS), and rates of nonalcoholic fatty liver disease (NAFLD) in the United States. We examined the effect of different amounts and forms of dietary fructose on the incidence or prevalence of NAFLD and indexes of liver health in humans. We conducted a systematic review of English-language, human studies of any design in children and adults with low to no alcohol intake and that reported at least one predetermined measure of liver health. The strength of the evidence was evaluated by considering risk of bias, consistency, directness, and precision. Six observational studies and 21 intervention studies met the inclusion criteria. The overall strength of evidence for observational studies was rated insufficient because of high risk of biases and inconsistent study findings. Of 21 intervention studies, 19 studies were in adults without NAFLD (predominantly healthy, young men) and 1 study each in adults or children with NAFLD. We found a low level of evidence that a hypercaloric fructose diet (supplemented by pure fructose) increases liver fat and aspartate aminotransferase (AST) concentrations in healthy men compared with the consumption of a weight-maintenance diet. In addition, there was a low level of evidence that hypercaloric fructose and glucose diets have similar effects on liver fat and liver enzymes in healthy adults. There was insufficient evidence to draw a conclusion for effects of HFCS or sucrose on NAFLD. On the basis of indirect comparisons across study findings, the apparent association between indexes of liver health (ie, liver fat, hepatic de novo lipogenesis, alanine aminotransferase, AST, and γ-glutamyl transpeptase) and fructose or sucrose intake appear to be confounded by excessive energy intake. Overall, the available evidence is not sufficiently robust to draw conclusions regarding effects of fructose, HFCS, or sucrose consumption on NAFLD.


Selected measures of health status for Mexican-American, mainland Puerto Rican, and Cuban-American children.
The 1987 National Vital Statistics System and the Hispanic Health and Nutrition Examination Survey (1982 through 1984) were used to assess the health status of Mexican-American, mainland Puerto Rican, and Cuban-American children by examining the prevalences of pregnancy outcomes and chronic medical conditions. The low-birth-weight rate among Hispanics (7.0%) compared favorably with that of non-Hispanic whites (7.1%) despite the greater poverty and lower levels of education among Hispanics. When examined by Hispanic subgroup, however, significant differences were present, with mainland Puerto Ricans having the highest prevalences of low-birth-weight infants. Premature births were more common among all three Hispanic subgroups than among non-Hispanic whites. Mexican-American and Cuban-American children had a similar prevalence of (3.9% and 2.5%, respectively) chronic medical conditions compared with non-Hispanic white children; Puerto Rican children had a higher prevalence of chronic medical conditions (6.2%). When assessed by these health status indicators, Hispanic children seem to have a health status similar to non-Hispanic white children. However, mainland Puerto Rican children seem at greater risk for poor health, reflecting the US Hispanic population's heterogeneity. Health programs targeted at US Hispanics should appropriately consider these group differences.


Low haemosporidian diversity and one key-host species in a bird malaria community on a mid-Atlantic island (São Miguel, Azores).
When host species colonize new areas, the parasite assemblage infecting the hosts might change, with some parasite species being lost and others newly acquired. These changes would likely lead to novel selective forces on both host and its parasites. We investigated the avian blood parasites in the passerine bird community on the mid-Atlantic island of São Miguel, Azores, a bird community originating from continental Europe. The presence of haemosporidian blood parasites belonging to the genera Haemoproteus, Plasmodium, and Leucocytozoon was assessed using polymerase chain reaction. We found two Plasmodium lineages and two Leucocytozoon lineages in 11 bird species (84% of all breeding passerine species) on the island. These lineages were unevenly distributed across bird species. The Eurasian Blackbird (Turdus merula) was the key-host species (total parasite prevalence of 57%), harboring the main proportion of parasite infections. Except for Eurasian Blackbirds, all bird species had significantly lower prevalence and parasite diversity compared to their continental populations. We propose that in evolutionary novel bird communities, single species may act as key hosts by harboring the main part of the parasite fauna from which parasites "leak" into the other species. This would create very different host-parasite associations in areas recently colonized by hosts as compared to in their source populations.


Venous thromboembolism after trauma: a never event?*.
Rates of venous thromboembolism as high as 58% have been reported after trauma, but there is no widely accepted screening protocol. If Medicare adds venous thromboembolism to the list of "preventable complications," they will no longer reimburse for treatment, which could have devastating effects on many urban centers. We hypothesized that prescreening with a risk assessment profile followed by routine surveillance with venous duplex ultrasound that could identify asymptomatic venous thromboembolism in trauma patients. Prospective, observational trial with waiver of consent. Level I trauma center intensive care unit. At admission, 534 patients were prescreened with a risk assessment profile. Patients (n = 106) with risk assessment profile scores >10 were considered high risk and received routine screening venous duplex ultrasound within 24 hrs and weekly thereafter. In prescreened high-risk patients, 20 asymptomatic deep vein thrombosis were detected with venous duplex ultrasound (19%). An additional ten venous thromboembolisms occurred, including six symptomatic deep vein thrombosis and four pulmonary emboli, resulting in an overall venous thromboembolism rate of 28%. The most common risk factors discriminating venous thromboembolism vs. no venous thromboembolism were femoral central venous catheter (23% vs. 8%), operative intervention >2 hrs (77% vs. 46%), complex lower extremity fracture (53% vs. 32%), and pelvic fracture (70% vs. 47%), respectively (all p < .05). Risk assessment profile scores were higher in patients with venous thromboembolism (19 ± 6 vs. 14 ± 4, p = .001). Risk assessment profile score (odds ratio 1.14) and the combination of pelvic fracture requiring operative intervention >2 hrs (odds ratio 5.75) were independent predictors for development of venous thromboembolism. The rates of venous thromboembolism for no chemical prophylaxis (33%), unfractionated heparin (29%), dalteparin (40%), or inferior vena cava filters (20%) were not statistically different (p = .764). Medicare's inclusion of venous thromboembolism after trauma as a "never event" should be questioned. In trauma patients, high-risk assessment profile score and pelvic fracture with prolonged operative intervention are independent predictors for venous thromboembolism development, despite thromboprophylaxis. Although routine venous duplex ultrasound screening may not be cost-effective for all trauma patients, prescreening using risk assessment profile yielded a cohort of patients with a high prevalence of venous thromboembolism. In such high-risk patients, routine venous duplex ultrasound and/or more aggressive prophylactic regimens may be beneficial.


Effect of the high-pitch mode in dual-source computed tomography on the accuracy of three-dimensional volumetry of solid pulmonary nodules: a phantom study.
To evaluate the influence of high-pitch mode (HPM) in dual-source computed tomography (DSCT) on the accuracy of three-dimensional (3D) volumetry for solid pulmonary nodules. A lung phantom implanted with 45 solid pulmonary nodules (n = 15 for each of 4-mm, 6-mm, and 8-mm in diameter) was scanned twice, first in conventional pitch mode (CPM) and then in HPM using DSCT. The relative percentage volume errors (RPEs) of 3D volumetry were compared between the HPM and CPM. In addition, the intermode volume variability (IVV) of 3D volumetry was calculated. In the measurement of the 6-mm and 8-mm nodules, there was no significant difference in RPE (p > 0.05, respectively) between the CPM and HPM (IVVs of 1.2 ± 0.9%, and 1.7 ± 1.5%, respectively). In the measurement of the 4-mm nodules, the mean RPE in the HPM (35.1 ± 7.4%) was significantly greater (p < 0.01) than that in the CPM (18.4 ± 5.3%), with an IVV of 13.1 ± 6.6%. However, the IVVs were in an acceptable range (< 25%), regardless of nodule size. The accuracy of 3D volumetry with HPM for solid pulmonary nodule is comparable to that with CPM. However, the use of HPM may adversely affect the accuracy of 3D volumetry for smaller (< 5 mm in diameter) nodule.


Rich club organization supports a diverse set of functional network configurations.
Brain function relies on the flexible integration of a diverse set of segregated cortical modules, with the structural connectivity of the brain being a fundamentally important factor in shaping the brain's functional dynamics. Following up on macroscopic studies showing the existence of centrally connected nodes in the mammalian brain, combined with the notion that these putative brain hubs may form a dense interconnected 'rich club' collective, we hypothesized that brain connectivity might involve a rich club type of architecture to promote a repertoire of different and flexibly accessible brain functions. With the rich club suggested to play an important role in global brain communication, examining the effects of a rich club organization on the functional repertoire of physical systems in general, and the brain in particular, is of keen interest. Here we elucidate these effects using a spin glass model of neural networks for simulating stable configurations of cortical activity. Using simulations, we show that the presence of a rich club increases the set of attractors and hence the diversity of the functional repertoire over and above the effects produced by scale free type topology alone. Within the networks' overall functional repertoire rich nodes are shown to be important for enabling a high level of dynamic integrations of low-degree nodes to form functional networks. This suggests that the rich club serves as an important backbone for numerous co-activation patterns among peripheral nodes of the network. In addition, applying the spin glass model to empirical anatomical data of the human brain, we show that the positive effects on the functional repertoire attributed to the rich club phenomenon can be observed for the brain as well. We conclude that a rich club organization in network architectures may be crucial for the facilitation and integration of a diverse number of segregated functions.


Lymphotoxin α fine-tunes T cell clonal deletion by regulating thymic entry of antigen-presenting cells.
Medullary thymic epithelial cells (mTEC) purge the T cell repertoire of autoreactive thymocytes. Although dendritic cells (DC) reinforce this process by transporting innocuous peripheral self-antigens, the mechanisms that control their thymic entry remain unclear. Here we show that mTEC-CD4+ thymocyte crosstalk regulates the thymus homing of SHPS-1+ conventional DCs (cDC), plasmacytoid DCs (pDC) and macrophages. This homing process is controlled by lymphotoxin α (LTα), which negatively regulates CCL2, CCL8 and CCL12 chemokines in mTECs. Consequently, Ltα-deficient mice have increased expression of these chemokines that correlates with augmented classical NF-κB subunits and increased thymic recruitment of cDCs, pDCs and macrophages. This enhanced migration depends mainly on the chemokine receptor CCR2, and increases thymic clonal deletion. Altogether, this study identifies a fine-tuning mechanism of T cell repertoire selection and paves the way for therapeutic interventions to treat autoimmune disorders.


OGT Regulates Mitochondrial Biogenesis and Function via Diabetes Susceptibility Gene Pdx1.
O-GlcNAc transferase (OGT), a nutrient sensor sensitive to glucose flux, is highly expressed in the pancreas. However, the role of OGT in the mitochondria of β-cells is unexplored. In this study, we identified the role of OGT in mitochondrial function in β-cells. Constitutive deletion of OGT (βOGTKO) or inducible ablation in mature β-cells (iβOGTKO) causes distinct effects on mitochondrial morphology and function. Islets from βOGTKO, but not iβOGTKO, mice display swollen mitochondria, reduced glucose-stimulated oxygen consumption rate, ATP production, and glycolysis. Alleviating endoplasmic reticulum stress by genetic deletion of Chop did not rescue the mitochondrial dysfunction in βOGTKO mice. We identified altered islet proteome between βOGTKO and iβOGTKO mice. Pancreatic and duodenal homeobox 1 (Pdx1) was reduced in in βOGTKO islets. Pdx1 overexpression increased insulin content and improved mitochondrial morphology and function in βOGTKO islets. These data underscore the essential role of OGT in regulating β-cell mitochondrial morphology and bioenergetics. In conclusion, OGT couples nutrient signal and mitochondrial function to promote normal β-cell physiology.


Terminal differentiation of osteoblasts to osteocytes is accompanied by dramatic changes in the distribution of actin-binding proteins.
Immunofluorescence staining of actin-binding proteins in osteoblasts and osteocytes was performed. alpha-Actinin, myosin, and tropomyosin showed similar organization in both osteoblastic stress fibers and osteocyte processes. However, fimbrin, villin, filamin, and spectrin showed dramatic differences in distribution between osteoblasts and osteocytes. This study suggested that terminal differentiation of osteoblasts to osteocytes is accompanied by highly dramatic changes in the distribution of actin-binding proteins. We previously reported that osteocyte shape is dependent on actin filaments. To analyze the terminal differentiation from osteoblasts to osteocytes, we investigated the actin-binding proteins, which are the control elements in the dynamic organization of the actin cytoskeleton. We used primary chick osteocytes and osteoblasts, the phenotypes of which were confirmed by use of OB7.3, a chick osteocyte-specific monoclonal antibody and by detection of alkaline phosphatase activity, respectively. Immunofluorescence staining was performed for visualizing actin-binding proteins. Furthermore, we applied shear stress at 12 dyns/cm2 to the cells and compared the changes in fimbrin distribution. Immunofluorescence staining of fimbrin and alpha-actinin showed their presence in the processes of osteocytes, with especially strong signals of fimbrin at the sites of divarication of the processes. Anti-villin was reactive with the osteocyte cytoplasm but not with the processes. Interestingly, anti-villin immunoreactivity was much stronger in osteocytes than in osteoblasts. Filamin was localized along the stress fibers of osteoblasts but was seen only in those in the proximal base of osteocyte processes. Myosin and tropomyosin were found to have a similar pattern in both stress fibers of osteoblasts and osteocyte processes. The difference in the distribution of anti-spectrin staining was highly dramatic. Osteoblasts immunostained with anti-spectrin showed punctate signals on their cytoplasmic membranes, whereas anti-spectrin in osteocytes detected a filamentous organization; and the spectrin was totally colocalized with actin from the distal portion of the cytoplasmic processes to the cell center. In osteoblasts, shear stress induced recruitment of fimbrin to the end of stress fibers. However, fimbrin in the osteocyte processes did not change its localization. We found that terminal differentiation of osteoblasts to osteocytes was accompanied by highly dramatic changes in the distribution of actin-binding proteins, changes of which may affect cellular function.


Ultrasonography and computed tomography in severe urinary tract infection.
A prospective study evaluated the utility of renal computed tomography (CT) and ultrasonography in 35 patients hospitalized for treatment of urinary tract infection. Renal computed tomograms were abnormal in 18 of 28 patients with acute pyelonephritis and three of four patients with urosepsis, showing findings consistent with pyelonephritis in 17 patients and intrarenal abscess or focal bacterial nephritis in four patients. Renal sonograms were abnormal in only eight patients, showing findings compatible with pyelonephritis in four and intrarenal abscess or focal bacterial nephritis in the other four. Flank tenderness was absent in only four patients with CT findings of pyelonephritis, of whom three were diabetic. We therefore found that (1) renal CT is a sensitive test for acute upper urinary tract infection, (2) ultrasonography detects focal bacterial nephritis and abscesses but is insensitive to uncomplicated upper urinary tract infection, and (3) painless pyelonephritis may be more common in patients with diabetes mellitus.


DUSP5 expression associates with poor prognosis in human neuroblastoma.
Regulation of growth and differentiation of neuroblastoma (NB) cells is the rational of some maintenance therapies for high-risk NB. MAP kinase phosphatases (MKPs) are potential physiologic regulators of neuronal differentiation and survival, but their expression patterns in NB are scarcely known. Here, an expression analysis of the MKP family has been performed using human NB tumor samples and human NB cell lines (SH-SY5Y, SMS-KCNR, and IMR-32) undergoing retinoic acid (RA)-induced differentiation or subjected to stimuli that activate the MAPK ERK1/2 pathway. We have identified candidate MKPs that could modulate differentiation and growth of NB cells. pERK1/2 high expression correlated with high expression of the MKP DUSP5 in NB tumors, and was associated with poor prognosis. ERK1/2 activation on SH-SY5Y cells was accompanied by increased cell proliferation, and correlated with the expression levels of DUSP5. Accordingly, siRNA knock-down of DUSP5 augmented proliferation of SH-SY5Y cells. Our findings provide insights into the dynamic expression of MKPs in NB cells, disclose DUSP5 as a potential marker of NB poor prognosis, and suggest a role for DUSP5 in limiting ERK1/2-mediated NB proliferation.


Structurally distinct membrane nanotubes between human macrophages support long-distance vesicular traffic or surfing of bacteria.
We report that two classes of membrane nanotubes between human monocyte-derived macrophages can be distinguished by their cytoskeletal structure and their functional properties. Thin membrane nanotubes contained only F-actin, whereas thicker nanotubes, i.e., those > approximately 0.7 microm in diameter, contained both F-actin and microtubules. Bacteria could be trapped and surf along thin, but not thick, membrane nanotubes toward connected macrophage cell bodies. Once at the cell body, bacteria could then be phagocytosed. The movement of bacteria is aided by a constitutive flow of the nanotube surface because streptavidin-coated beads were similarly able to traffic along nanotubes between surface-biotinylated macrophages. Mitochondria and intracellular vesicles, including late endosomes and lysosomes, could be detected within thick, but not thin, membrane nanotubes. Analysis from kymographs demonstrated that vesicles moved in a stepwise, bidirectional manner at approximately 1 microm/s, consistent with their traffic being mediated by the microtubules found only in thick nanotubes. Vesicular traffic in thick nanotubes and surfing of beads along thin nanotubes were both stopped upon the addition of azide, demonstrating that both processes require ATP. However, microtubule destabilizing agents colchicine or nocodazole abrogated vesicular transport but not the flow of the nanotube surface, confirming that distinct cytoskeletal structures of nanotubes give rise to different functional properties. Thus, membrane nanotubes between macrophages are more complex than unvarying ubiquitous membrane tethers and facilitate several means for distal interactions between immune cells.


Terminal loop-mediated control of microRNA biogenesis.
Regulation of miRNA (microRNA) biogenesis shapes the profiles of miRNAs in the living cell, contributing to cell identity and function. Importantly, aberrant miRNA levels have been linked to a variety of human pathological states. In recent years, a number of proteins have been shown to regulate the miRNA biogenesis at the level of Drosha and Dicer cleavage. A large proportion of these factors regulate miRNA production through binding to the TL (terminal loop) regions of miRNA progenitors. In the present paper, we review the current knowledge about pri-miRNA (primary miRNA) and pre-miRNA (precursor miRNA) TL involvement in the regulation of miRNA biogenesis.


Cell fission and formation of mini cell bodies by high frequency alternating electric field.
We report the use of high frequency alternating electric fields (AC) to induce deformation of sea urchin eggs, leading to budding of membrane vesicles or fission of cells. Several mini cell bodies can be prepared from a single egg by carefully manipulating the frequency and amplitude of the AC field and the ratio between the interelectrode spacing and the cell diameter, alpha. alpha values between 2.2 and 3.5 have been found to be optimal for inducing fission of sea urchin eggs. In a typical experiment, a sea urchin egg (diameter = 75 microns), suspended in a low ionic medium (conductance < 2 mS/m), was located under the microscope between two platinum wire electrodes, separated by a distance of approximately 200 microns. A medium strength AC field (< 100 V/cm at 2 MHz) was applied to attract the egg to one of the two electrodes via dielectrophoresis. This process took place in a few seconds. The voltage was then slowly increased to approximately 1000 V/cm over approximately 30 s. The cell elongated and separated into two fragments, the larger one containing the nucleus. When the field was turned off, the mother cell and the daughter vesicle retracted to form spherical mini cell bodies that appear to be stable as assessed by the absence of swelling for the duration of the experiment (approximately 15 min). This indicates that membranes of these mini cell bodies were not leaky to ions and small molecules. This procedure could be repeated a few times to make several mini cell bodies from a single egg. With practice, several minicell bodies could also be prepared in a single fission experiment by adjusting the field parameters and the a value. Cell fission is a result of the mechanical stress produced by the AC field. These procedures may be used to prepare mini membrane vesicles for voltage clamp experiments or to perform microsurgical manipulation of cells, embryos, or chromosomes.


Allorecognition mechanisms during ascidian fertilization.
Ascidians (primitive chordates) are hermaphroditic animals, releasing sperm and eggs nearly simultaneously. But, many ascidians, including Ciona intestinalis and Halocynthia roretzi, show self-sterility or preference for cross-fertilization rather than self-fertilization. The molecular mechanisms underlying this allorecognition process are only poorly understood. We recently identified the genes responsible for self-incompatibility in C. intestinalis by a positional cloning: sperm-borne polycystin 1-like receptor, referred to as s-Themis, and its fibrinogen-like ligand called v-Themis on the vitelline coat (VC) are highly polymorphic and appear to be responsible for allorecognition in the fertilization of C. intestinalis. In H. roretzi, on the other hand, we revealed that HrVC70, a 70-kDa main component of the VC consisting of 12 epidermal-growth-factor (EGF)-like repeats, is a candidate allorecognition protein, since the attachment of this protein to the VC during oocyte maturation and its detachment by weak acid are closely linked to the gain and the loss of self-sterility, respectively, and also since nonself-sperm rather than self-sperm efficiently bound to HrVC70-agarose. As a binding partner of HrVC70, a 35-kDa GPI-anchored glycoprotein in sperm lipid rafts, referred to as HrUrabin, was identified: HrUrabin appears to play a key role in allorecognizable sperm binding to HrVC70 during fertilization. In the present review, we describe the current progress on the molecular bases of allorecognition, or self-incompatibility, during ascidian fertilization, by considering the SI systems in another organisms including fungies and flowering plants.


Downregulation of Msx-2 expression results in chondrogenesis in the medial region of the avian mandible.
Homeobox-containing genes are thought to be regulators of pattern formation during vertebrate development. We have previously characterized regions in the developing chick mandible expressing either Msx-1 or Msx-2 in terms of their chondrogenic potential, rates of cell proliferation, and their correlation with regions of cell death. These experiments suggest that Msx-1 may be involved in outgrowth of the mandibular arch, and Msx-2 may be involved in delineating the non-chondrogenic region of the medial part of the mandibular arch. To further examine the possibility that expression of Msx-2 may be involved in preventing chondrogenesis in the medial region, mandibular arch explants from stage 23 chick embryos were cultured for 4 days in media in the absence of serum but in the presence of 20-30 microM Msx-2 sense or antisense oligonucleotides (18 mers). In explants grown in either control media or with the sense oligonucleotide two rods of cartilage separated by a cartilage free area located in the medial region of the mandible were formed. In explants treated with Msx-2 antisense oligonucleotide cartilage formation was observed in the medial region of the mandible resulting in the fusion of the two bilateral rods at the midline. These results suggest a negative relationship between Msx-2 expression and chondrogenesis in the medial region of the developing mandible.


Erythropoietin inhalation enhances adult canine alveolar-capillary formation following pneumonectomy.
Paracrine erythropoietin (EPO) signaling in the lung recruits endothelial progenitor cells, promotes cell maturation and angiogenesis, and is upregulated during canine postpneumonectomy (PNX) compensatory lung growth. To determine whether inhalational delivery of exogenous EPO augments endogenous post-PNX lung growth, adult canines underwent right PNX and received, via a permanent tracheal stoma, weekly nebulization of recombinant human EPO-containing nanoparticles or empty nanoparticles (control) for 16 wk. Lung function was assessed under anesthesia pre- and post-PNX. The remaining lobes were fixed for detailed morphometric analysis. Compared with control treatment, EPO delivery significantly increased serum EPO concentration without altering systemic hematocrit or hemoglobin concentration and abrogated post-PNX lipid oxidative stress damage. EPO delivery modestly increased post-PNX volume densities of the alveolar septum per unit of lung volume and type II epithelium and endothelium per unit of septal tissue volume in selected lobes. EPO delivery also augmented the post-PNX increase in alveolar double-capillary profiles, a marker of intussusceptive capillary formation, in all remaining lobes. EPO treatment did not significantly alter absolute resting lung volumes, lung and membrane diffusing capacities, alveolar-capillary blood volume, pulmonary blood flow, lung compliance, or extravascular alveolar tissue volumes or surface areas. Results established the feasibility of chronic inhalational delivery of growth-modifying biologics in a large animal model. Exogenous EPO selectively enhanced cytoprotection and alveolar angiogenesis in remaining lobes but not whole-lung extravascular tissue growth or resting function; the nonuniform response contributes to structure-function discrepancy, a major challenge for interventions aimed at amplifying the innate potential for compensatory lung growth.


Human subcortical brain asymmetries in 15,847 people worldwide reveal effects of age and sex.
The two hemispheres of the human brain differ functionally and structurally. Despite over a century of research, the extent to which brain asymmetry is influenced by sex, handedness, age, and genetic factors is still controversial. Here we present the largest ever analysis of subcortical brain asymmetries, in a harmonized multi-site study using meta-analysis methods. Volumetric asymmetry of seven subcortical structures was assessed in 15,847 MRI scans from 52 datasets worldwide. There were sex differences in the asymmetry of the globus pallidus and putamen. Heritability estimates, derived from 1170 subjects belonging to 71 extended pedigrees, revealed that additive genetic factors influenced the asymmetry of these two structures and that of the hippocampus and thalamus. Handedness had no detectable effect on subcortical asymmetries, even in this unprecedented sample size, but the asymmetry of the putamen varied with age. Genetic drivers of asymmetry in the hippocampus, thalamus and basal ganglia may affect variability in human cognition, including susceptibility to psychiatric disorders.


Popular belief meets surgical reality: impact of lunar phases, Friday the 13th and zodiac signs on emergency operations and intraoperative blood loss.
The influence of superstition, moon calendars, and popular belief on evidence-based medicine is stunning. More than 40% of medical staff is convinced that lunar phases can affect human behavior. The idea that Friday the 13th is associated with adverse events and bad luck is deep-rooted in the population of Western industrial countries. The aim of the present study was to test the hypothesis that these myths are transferable to real-life surgery. We analyzed the extent to which moon phases, zodiac signs, and Friday the 13th influence blood loss, emergency frequency, and intestinal perforations by evaluating the operation records of all 27,914 consecutive patients of our institution undergoing general, visceral, or vascular surgery between August 2001 and August 2010. Dates of surgery were allocated to lunar phases and to zodiac signs, as well as to Friday the 13th. A total of 111 lunar cycles and 15 Fridays the 13th occurred within the 3,281-day observation period. Patients' characteristics did not differ in lunar phases, zodiac signs, or Fridays the 13th. Full moon phases, the presence of Friday the 13th, and zodiac signs influenced neither intraoperative blood loss nor emergency frequency. No statistical peaks regarding perforated aortic aneurysms and gastrointestinal perforations were found on full moon or Friday the 13th. Scientific analysis of our data does not support the belief that moon phases, zodiac signs, or Friday 13th influence surgical blood loss and emergency frequency. Our data indicate that such beliefs are myths far beyond reality.


Aurora kinase A as a rational target for therapy in glioblastoma.
Despite advances in the knowledge of tumor biology, the outcome of glioblastoma tumors remains poor. The design of many molecularly targeted therapies in glioblastoma has focused on inhibiting molecular abnormalities present in tumor cells compared with normal tissue rather than patient outcome-associated factors. As an alternative approach, the present study identified genes associated with shorter survival as potential therapeutic targets. It was hypothesized that inhibition of a molecular target associated with poor outcome would impact glioblastoma cell proliferation. The present study correlated patient survival data with tumor gene expression profiling and gene ontology analysis. Genes associated with shorter survival were identified and one of these was selected for therapeutic targeting in an in vitro system. Glioblastoma cell growth suppression was measured by H(3)-thymidine uptake, colony formation, and flow cytometry. The gene expression microarray and ontology analysis revealed that genes involved in mitotic processes, including AURKA, were associated with poor prognosis in glioblastoma. Inhibition of AURKA suppressed glioblastoma cell growth. Moreover, inhibition of AURKA was synergistic with radiation in glioblastoma cells at high radiation doses. Relative expression of AURKA may be of prognostic value and warrants further investigation with larger, prospective studies. Pharmacological inhibition of AURKA is a potentially promising therapy for glioblastoma.


Delayed presentation of late-onset cerebrospinal fluid rhinorrhoea following dopamine agonist therapy for giant prolactinoma.
Therapeutic shrinkage of prolactinomas with dopamine agonists achieves clinical benefit but can expose fistulae that have arisen as a result of bony erosion of the sella floor and anterior skull base by the invasive tumour, resulting in the potential development of cerebrospinal fluid (CSF) rhinorrhoea, meningitis, and rarely pneumocephalus. Onset of symptoms is typically within 4 months of commencing therapy. The management is typically surgical repair via an endoscopic transnasal transsphenoidal approach. A 23-year-old man presented to the Emergency Department with acute left limb weakness and intermittent headaches. Visual fields were full to confrontation. Immediate computed tomography and subsequent magnetic resonance imaging (MRI), demonstrated a 5 cm lobular/cystic mass invading the right cavernous sinus, displacing and compressing the midbrain, with destruction of the bony sella. He was referred to the regional pituitary multidisciplinary team (MDT). Serum prolactin was 159 455 mIU/l (7514.37 ng/ml) (normal ranges 100-410 mIU/l (4.72-19.34 ng/ml)). Cabergoline was commenced causing dramatic reduction in tumour size and resolution of neurological symptoms. Further dose titrations were required as the prolactin level plateaued and significant residual tumour remained. After 13 months of treatment, he developed continuous daily rhinorrhea, and on presenting to his general practitioner was referred to an otolaryngologist. When next seen in the routine regional pituitary clinic six-months later he was admitted for urgent surgical repair. Histology confirmed a prolactinoma with a low proliferation index of 2% (Ki-67 antibody). In view of partial cabergoline resistance he completed a course of conventional radiotherapy. Nine months after treatment the serum prolactin had fallen to 621 mIU/l, and 12 months after an MRI showed reduced tumour volume. CSF rhinorrhoea occurred 13 months after the initiation of cabergoline, suggesting a need for vigilance throughout therapy.Dedicated bony imaging should be reviewed early in the patient pathway to assess the potential risk of CSF rhinorrhoea after initiation of dopamine agonist therapy.There was a significant delay before this complication was brought to the attention of the regional pituitary MDT, with associated risk whilst left untreated. This demonstrates a need for patients and healthcare professionals to be educated about early recognition and management of this complication to facilitate timely and appropriate referral to the MDT for specialist advice and management. We changed our nurse-led patient education programme as a result of this case.An excellent therapeutic response was achieved with conventional radiotherapy after limited surgery having developed partial cabergoline resistance and CSF rhinorrhoea.


A review on biochar modulated soil condition improvements and nutrient dynamics concerning crop yields: Pathways to climate change mitigation and global food security.
The beneficial role of biochar on improvement of soil quality, C sequestration, and enhancing crop yield is widely reported. As such there is not much consolidated information available linking biochar modulated soil condition improvement and soil nutrient availability on crop yields. The present review paper addresses the above issues by compilation of world literature on biochar and a new dimension is introduced in this review by performing a meta-analysis of published data by using multivariate statistical analysis. Hence this review is a new in its kind and is useful to the broad spectrum of readers. Generally, alkalinity in biochar increases with increase in pyrolysis temperature and majority of the biochar is alkaline in nature except a few which are acidic. The N content in many biochar was reported to be more than 4% as well as less than 0.5%. Poultry litter biochar is a rich source of P (3.12%) and K (7.40%), while paper mill sludge biochar is higher in Ca content (31.1%) and swine solids biochar in Zn (49810 mg kg-1), and Fe (74800 mg kg-1) contents. The effect of biochar on enhancing soil pH was higher in Alfisol, Ferrosol and Acrisol. Soil application of biochar could on an average increase (78%), decrease (16%), or show no effect on crop yields under different soil types. Biochar produced at a lower pyrolysis temperature could deliver greater soil nutrient availabilities than that prepared at higher temperature. Principal component analysis (PCA) of available data shows an inverse relationship between [pyrolysis temperature and soil pH], and [biochar application rate and soil cation exchange capacity]. The PCA also suggests that the original soil properties and application rate strongly control crop yield stimulations via biochar amendments. Finally, biochar application shows net soil C gains while also serving for increased plant biomass production that strongly recommends biochar as a useful soil amendment. Therefore, the application of biochar to soils emerges as a 'win-win strategy' for sustainable waste management, climate change mitigation and food security.


Long-term sequelae of conservative treatment by surgery, brachytherapy, and chemotherapy for vulval and vaginal rhabdomyosarcoma in children.
Between 1970 and 1978, 17 girls with rhabdomyosarcoma (RMS) of the vulva or vagina were treated at the Institut Gustave-Roussy (IGR) by conservative treatment including surgery, brachytherapy, and chemotherapy. Twelve pubescent or postpubescent girls were studied for long-term sequelae. Eleven of 12 patients have had a normal puberty, two have a total of three healthy children, 11 have normal menses, and 10 normal menarche (one after hormonal replacement). Only one patient underwent hysterectomy following low-dose brachytherapy. Five girls have no vaginal complications, and three are sexually active. Three have had minimal vaginal sequelae, which required surgical correction to permit sexual intercourse. Four girls sustained serious sequelae (colorectal, vaginal, urethral, and ureteral stenosis). These sequelae, secondary to irradiation, are potentially avoidable in the future given the current advances in brachytherapy and improvements in dosimetry. This conservative treatment is useful and appropriate for girls with RMS of vulvar or vaginal origin when complete remission cannot be obtained with chemotherapy and partial colpectomy.


High susceptibility of MDR and XDR Gram-negative pathogens to biphenyl-diacetylene-based difluoromethyl-allo-threonyl-hydroxamate LpxC inhibitors.
Inhibitors of uridine diphosphate-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine deacetylase (LpxC, which catalyses the first, irreversible step in lipid A biosynthesis) are a promising new class of antibiotics against Gram-negative bacteria. The objectives of the present study were to: (i) compare the antibiotic activities of three LpxC inhibitors (LPC-058, LPC-011 and LPC-087) and the reference inhibitor CHIR-090 against Gram-negative bacilli (including MDR and XDR isolates); and (ii) investigate the effect of combining these inhibitors with conventional antibiotics. MICs were determined for 369 clinical isolates (234 Enterobacteriaceae and 135 non-fermentative Gram-negative bacilli). Time-kill assays with LPC-058 were performed on four MDR/XDR strains, including Escherichia coli producing CTX-M-15 ESBL and Klebsiella pneumoniae, Pseudomonas aeruginosa and Acinetobacter baumannii producing KPC-2, VIM-1 and OXA-23 carbapenemases, respectively. LPC-058 was the most potent antibiotic and displayed the broadest spectrum of antimicrobial activity, with MIC90 values for Enterobacteriaceae, P. aeruginosa, Burkholderia cepacia and A. baumannii of 0.12, 0.5, 1 and 1 mg/L, respectively. LPC-058 was bactericidal at 1× or 2× MIC against CTX-M-15, KPC-2 and VIM-1 carbapenemase-producing strains and bacteriostatic at ≤4× MIC against OXA-23 carbapenemase-producing A. baumannii. Combinations of LPC-058 with β-lactams, amikacin and ciprofloxacin were synergistic against these strains, albeit in a species-dependent manner. LPC-058's high efficacy was attributed to the presence of the difluoromethyl-allo-threonyl head group and a linear biphenyl-diacetylene tail group. These in vitro data highlight the therapeutic potential of the new LpxC inhibitor LPC-058 against MDR/XDR strains and set the stage for subsequent in vivo studies.


On-line simultaneous monitoring of polarization mode dispersion and chromatic dispersion.
We have developed new expressions for power fading and average power fading owing to polarization mode dispersion (PMD) and chromatic dispersion in terms of the angle of precession of the output state of polarization about the PMD vector. Based on these expressions, a simple and novel pilot-tone-based technique for simultaneous monitoring of chromatic dispersion and PMD is proposed. Experimental results demonstrate the effectiveness of the proposed technique; these results agree well with the theoretical analysis.


The versatile functions of Sox9 in development, stem cells, and human diseases.
The transcription factor Sox9 was first discovered in patients with campomelic dysplasia, a haploinsufficiency disorder with skeletal deformities caused by dysregulation of Sox9 expression during chondrogenesis. Since then, its role as a cell fate determiner during embryonic development has been well characterized; Sox9 expression differentiates cells derived from all three germ layers into a large variety of specialized tissues and organs. However, recent data has shown that ectoderm- and endoderm-derived tissues continue to express Sox9 in mature organs and stem cell pools, suggesting its role in cell maintenance and specification during adult life. The versatility of Sox9 may be explained by a combination of post-transcriptional modifications, binding partners, and the tissue type in which it is expressed. Considering its importance during both development and adult life, it follows that dysregulation of Sox9 has been implicated in various congenital and acquired diseases, including fibrosis and cancer. This review provides a summary of the various roles of Sox9 in cell fate specification, stem cell biology, and related human diseases. Ultimately, understanding the mechanisms that regulate Sox9 will be crucial for developing effective therapies to treat disease caused by stem cell dysregulation or even reverse organ damage.


Cellular dynamics of rats' submaxillary gland during heat acclimatization.
Male laboratory rats (Rattus norvegicus) ranging between 130 and 160 g were subjected to ambient temperatures of 34.5 degrees C from 0 to 28 days. The submaxillary gland, liver, spleen, and kidney were weighed. The acini size and the mitotic index of the acinar cells after intraperitoneal injection of vinblastine sulfate and colchicine were recorded for the submaxillary and parotid glands. The submaxillary glands showed increased growth rate over the initial 5 days, whereas the growth rate of the other organs decreased. The submaxillary gland enlargement was due to an initial hyperplasia of the acini cells reaching a maximum on the 2nd day. This was followed by hypertrophy of the Acini reaching a maximum on the l0th day. The parotid acini at control values throughout the experiment. It is postulated that the gland enlargement is effected to increase evaporative heat loss during the early stages of acclimatization prior to the lowering of heat production resulting from biochemical adaptation.


The Clinica del Lavoro, the first clinic for occupational diseases of the world, was inaugurated in Milan on 20 March 1910; its first director was Luigi Devoto, who was in charge until 1935. The purpose of this work is to review the activities of industrial hygiene and toxicology carried out at the Clinica del Lavoro under the guidance of Devoto. Documents published by the Istituti Clinici di Perfezionamento, a group of clinics of which the Clinica del Lavoro was part, record the birth and organization of this structure and the presence of a laboratory of chemistry; documents by Devoto and other authors were also retrieved to extrapolate specific information on activities of industrial hygiene and toxicology. The Clinica del Lavoro, at the time of its inauguration, included four laboratories: of chemistry, clinical physics, histopathology and bacteriology. The chemistry lab was located on the first floor and was composed of 6 well-lit rooms, modernly equipped with work benches that could accommodate 12 people. In Devoto's view, the chemistry laboratory, supported by that of clinical physics, had to assess the toxicological properties of chemicals commonly found in the workplace and to reveal the mechanisms of induction of damage to humans. In the first 30 years of activity, the Clinica del Lavoro investigated various diseases deriving from exposure to chemical agents, including saturnism, or lead intoxication, mercurialism, phosphorism, benzolism, sulfocarbonism, dust diseases. Several assays were developed and applied to measure toxicants in different biological and environmental mean as evidenced by scientific publications starting from 1920. In Devoto's view, industrial hygiene and toxicology were essential tools for the research and prevention of occupational diseases since the first years of activity of the Clinica del Lavoro.


Capturing Collaborative Challenges: Designing Complexity-Sensitive Theories of Change for Cross-Sector Partnerships.
Systems change requires complex interventions. Cross-sector partnerships (CSPs) face the daunting task of addressing complex societal problems by aligning different backgrounds, values, ideas and resources. A major challenge for CSPs is how to link the type of partnership to the intervention needed to drive change. Intervention strategies are thereby increasingly based on Theories of Change (ToCs). Applying ToCs is often a donor requirement, but it also reflects the ambition of a partnership to enhance its transformative potential. The current use of ToCs in partnering efforts varies greatly. There is a tendency for a linear and relatively simple use of ToCs that does limited justice to the complexity of the problems partnerships aim to address. Since partnership dynamics are already complex and challenging themselves, confusion and disagreement over the appropriate application of ToCs is likely to hamper rather than enhance the transformative potential of partnerships. We develop a complexity alignment framework and a diagnostic tool that enables partnerships to better appreciate the complexity of the context in which they operate, allowing them to adjust their learning strategy. This paper applies recent insights into how to deal with complexity from both the evaluation and theory of change fields to studies investigating the transformative capacity of partnerships. This can (1) serve as a check to define the challenges of partnering projects and (2) can help delineate the societal sources and layers of complexity that cross-sector partnerships deal with such as failure, insufficient responsibility taking and collective action problems at four phases of partnering.


Incidence and natural history of proliferative sickle cell retinopathy: observations from a cohort study.
To describe the incidence, prevalence, and natural history of proliferative sickle cell retinopathy (PSR). Prospective longitudinal study over 20 years. Newborn screening of 100000 consecutive deliveries from 1973 to 1981 identified 315 children with homozygous sickle cell (SS) disease and 201 with SS-hemoglobin C (SC) disease. By the age of 5 years, 307 SS patients and 166 SC patients were alive and living in Jamaica and were recruited for this ophthalmic study. Description of retinal vascular changes on annual angiography and angioscopy. Incidence and prevalence of PSR and its behavior on follow-up. Progression of PSR was investigated using the number of eyes affected (none, one, both) and the interval until PSR onset. At last review in January 2000, PSR had developed in 59 patients (14 SS, 45 SC), unilaterally in 36 patients and bilaterally in 23. Incidence increased with age in both genotypes, with crude annual incidence rates of 0.5 cases (95% confidence interval [CI], 0.3-0.8) per 100 SS subjects and 2.5 cases (95% CI, 1.9-3.3) per 100 SC subjects. Prevalence was greater in SC disease, and by the ages of 24 to 26 years, PSR had occurred in 43% subjects with SC disease and in 14% subjects with SS disease. Patients with unilateral PSR had a 16% (11% SS, 17% SC) probability of regressing to no PSR and a 14% (16% SS, 13% SC) probability of progressing to bilateral PSR. Those with bilateral PSR had an 8% (8% SS, 8% SC) probability of regressing to unilateral PSR and a 1% (0 SS, 2% SC) probability of regressing to a PSR-free state. Irretrievable visual loss occurred in only 1 of 82 PSR-affected eyes, and 1 required detachment surgery and recovered normal visual acuity. Longitudinal observations over 20 years in a cohort of patients followed from birth confirms a greater incidence and severity of PSR in SC disease, and shows that spontaneous regression occurred in 32% of PSR-affected eyes. Permanent visual loss was uncommon in subjects observed up to the age of 26 years.


Single-incision videolaparoscopic appendectomy with conventional videolaparoscopy equipment.
Acute appendicitis is the most common surgical emergency in daily practice, and is approached laparoscopically in many centers. Efforts have been undertaken for the development of minimally invasive techniques that reduce tissue trauma and offer improved cosmetic results, one of such being the single-incision laparoscopic surgery (SILS). To present a minimally invasive technique for appendectomy (SILS) undertaken with conventional instruments. Eleven patients were treated in the emergency care center presenting abdominal pain in the right iliac fossa that was suggestive of appendicitis. Diagnostic investigation was subsequently conducted, including physical examination, laboratory and imaging exams (CT scan with intravenous contrast or total abdominal ultrasound), and the results were consistent with acute appendicitis. Thus, after consent, these patients underwent SILS appendectomy under general anesthesia with three trocars (two 10 mm and one 5 mm), using conventional and optical laparoscopic tweezers (10 mm, 30º). The base and pedicle of the appendix were ligated with titanium LT 400 clips. The procedure occurred uneventfully. Inclusion criteria were absence of diffuse peritonitis, BMI (body mass index) less than 35 and absence of serious comorbidities or sepsis. Seven men and four women were operated with average age of 25.7 years and underwent appendectomy through this technique. Mean procedure duration was of 37.2 min. Regarding surgical findings, three had appendicitis in stage 1, four in stage 2 and four in stage 3. All patients improved well, without surgical complications, and did not require conversion to open surgery or conventional laparoscopy technique. Appendectomy conducted through Single Incision Laparoscopic Surgery is a feasible and promising technique that can be performed with conventional laparoscopic instruments.


Role of nurse practitioners in reducing cardiovascular risk factors: a retrospective cohort study.
To determine the impact of nurse practitioners' counselling on reducing cardiovascular risk factors in patients participating in routine preventive check-ups. A new model of 'renewed' family practice was introduced in Slovenia as a pilot project in 2011, in which nurse practitioners are included in a team carrying out preventive activities and managing patients with stable chronic diseases. A retrospective cohort study. This study was conducted in 16 family medicine practices (eight renewed and eight regular family practices). In each family practice, a systematic sample was selected of registered patients participating in a cardiovascular preventive check-up. Data on sex, age, blood pressure, cholesterol, blood sugar, smoking, level of physical activity and cardiovascular risk were collected. Patients attending renewed family practices received counselling on risk factors from nurse practitioners (test group), and patients attending regular family practices received counselling from family physicians (control group). Data were collected again at least one and no more than five years after the baseline consultation. There were 128 patients in the test group and 129 patients in the control group. At the control visit, the patients counselled by nurse practitioners had significantly lower levels of systolic blood pressure and cholesterol and practiced regular physical activity significantly more often than patients counselled by family physicians. Nurse practitioners can be at least as successful as physicians when counselling patients on cardiovascular risk factors during their preventive check-ups. This study showed that nurse practitioners have an important role in managing patients at the primary care level.


Arrested development in Ancylostoma duodenale: course of a self-induced infection in man.
The course of self-induced infection with about 100 Ancylostoma duodenale larvae was followed by twice-weekly blood and stool examinations. After a small initial increase, no further change in eosinophil counts occurred until the 33rd week of infection when they began to increase sharply. Seven weeks later, eggs were first observed in stools. Thus, in this infection, the prepatent period was five times the expected. It is suggested that a strain of A.duodenale exists which either 1) has an inherent, abnormally long prepatent period or 2) has a prepatent period which is susceptible to change depending upon the environmental conditions experienced by the free-living larvae. In either case, the third stage larvae probably enter a hypobiotic state within the host which lasts for about 8 months.


Donor lymphocyte infusions modulate relapse risk in mixed chimeras and induce durable salvage in relapsed patients after T-cell-depleted allogeneic transplantation for Hodgkin's lymphoma.
Reduced-intensity conditioning has minimized nonrelapse-related mortality rates after allogeneic transplantation in patients with Hodgkin's lymphoma, and relapse has now become the major cause for treatment failure. We aimed to assess the impact of donor lymphocyte infusions (DLIs) on relapse incidence when administered for mixed chimerism and their utility as salvage therapy when given for relapse. This study reports the outcomes of 76 consecutive patients with multiply relapsed or refractory Hodgkin's lymphoma who underwent allogeneic transplantation that incorporated in vivo T-cell depletion. Forty-two patients had related donors and 34 had unrelated donors. DLIs were administered in a dose-escalating fashion to 22 patients for mixed chimerism (median time of first dose, 9 months post-transplantation) and to 24 patients for relapse. Three-year donor lymphocyte-related mortality was 7%, relating mainly to the induction of graft-versus-host disease. Nineteen (86%) of 22 patients receiving donor lymphocytes for mixed chimerism converted to full donor status. Four-year relapse incidence was 5% in these 22 patients compared with 43% in patients who remained relapse free but full donor chimeras at 9 months post-transplantation (P = .0071). Nineteen (79%) of 24 patients receiving donor lymphocytes for relapse responded (14 complete responses, five partial responses). Four-year overall survival from relapse was 59% in recipients of donor lymphocytes, contributing to a 4-year overall survival from transplantation of 64% and a 4-year current progression-free survival of 59% in all 76 patients. These data demonstrate the potential for allogeneic immunotherapy with donor lymphocytes both to reduce relapse risk and to induce durable antitumor responses in patients with Hodgkin's lymphoma after hematopoietic stem-cell transplantation that incorporates in vivo T-cell depletion.


Accounting for personal and professional choices for pandemic influenza vaccination amongst English healthcare workers.
Healthcare workers (HCWs) are encouraged to get vaccinated during influenza pandemics to reduce their own, and patients', risk of infection, and to encourage their patients to get immunised. Despite extensive research on HCWs' receipt of vaccination, little is known about how HCWs articulate pandemic influenza vaccination advice to patients. To explore HCWs' uptake of the A/H1N1 vaccine during the pandemic of 2009-2010, their recommendations to patients at the time, and their anticipated choices around influenza vaccination under different pandemic scenarios. We conducted semi-structured interviews and focus groups with eight vaccinated and seventeen non-vaccinated HCWs from primary care practices in England. The data was analysed using thematic analysis. The HCWs constructed their receipt of vaccination as a personal choice informed by personal health history and perceptions of vaccine safety, while they viewed patients' vaccination as choices made following informed consent and medical guidelines. Some HCWs received the A/H1N1 vaccine under the influence of their local practice organizational norms and values. While non-vaccinated HCWs regarded patients' vaccination as patients' choice, some vaccinated HCWs saw it also as a public health issue. The non-vaccinated HCWs emphasised that they would not allow their personal choices to influence the advice they gave to patients, whereas some vaccinated HCWs believed that by getting vaccinated themselves they could provide a reassuring example to patients, particularly those who have concerns about influenza vaccination. All HCWs indicated they would accept vaccination under the severe pandemic scenario. However, most non-vaccinated HCWs expressed reticence to vaccinate under the mild pandemic scenario. Providing evidence-based arguments about the safety of new vaccines and the priority of public health over personal choice, and creating strong social norms for influenza vaccination as part of the organizational culture, should increase uptake of influenza vaccination among primary care HCWs and their patients.